Michaelis-Menten Analysis of Alkaline Phosphatase by Capillary Electrophoresis Using Plug-Plug Reaction
作者:
Yan Xu,
Xuhui Liu,
MichaelP. C. Ip,
期刊:
Journal of Liquid Chromatography & Related Technologies
(Taylor Available online 1998)
卷期:
Volume 21,
issue 18
页码: 2781-2797
ISSN:1082-6076
年代: 1998
DOI:10.1080/10826079808003442
出版商: Taylor & Francis Group
数据来源: Taylor
摘要:
This work evaluated the use of an alkaline phosphatase plug, slowly migrating in a polyacrylamide-coated capillary filled with buffer/soluble polymer, to convertp-nitrophenylphosphate (which was injected into the capillary as a separate plug, and migrated faster than the enzyme) top-nitrophenol under a constant applied potential. The elution of assay components was monitored on-capillary at 230 nm. The initial reaction velocity of the enzymecatalyzed reaction was estimated from the peak area ratio of the enzyme product to the internal standard. Using the Lineweaver-Burk plots, an average Michaelis constant (KM) for alkaline phosphatase was calculated to be 4.8 ± 0.3 mM (n = 4, CV = 6%). With the constant potential electrophoresis (8 kV/57 cm), the method had a detection limit of 4.4 × 10−5IU for alkaline phosphatase and a linear range up to 2.1 × 10−3IU.
点击下载:
PDF (632KB)
返 回